Deegan Shane, Saveljeva Svetlana, Logue Susan E, Pakos-Zebrucka Karolina, Gupta Sanjeev, Vandenabeele Peter, Bertrand Mathieu J M, Samali Afshin
a Apoptosis Research Center; NUI Galway ; Galway , Ireland.
Autophagy. 2014;10(11):1921-36. doi: 10.4161/15548627.2014.981790.
Endoplasmic reticulum (ER) stress-induced cell death is normally associated with activation of the mitochondrial apoptotic pathway, which is characterized by CYCS (cytochrome c, somatic) release, apoptosome formation, and caspase activation, resulting in cell death. In this study, we demonstrate that under conditions of ER stress cells devoid of CASP9/caspase-9 or BAX and BAK1, and therefore defective in the mitochondrial apoptotic pathway, still undergo a delayed form of cell death associated with the activation of caspases, therefore revealing the existence of an alternative stress-induced caspase activation pathway. We identified CASP8/caspase-8 as the apical protease in this caspase cascade, and found that knockdown of either of the key autophagic genes, ATG5 or ATG7, impacted on CASP8 activation and cell death induction, highlighting the crucial role of autophagy in the activation of this novel ER stress-induced death pathway. In line with this, we identified a protein complex composed of ATG5, FADD, and pro-CASP8 whose assembly coincides with caspase activation and cell death induction. Together, our results reveal the toxic potential of autophagy in cells undergoing ER stress that are defective in the mitochondrial apoptotic pathway, and suggest a model in which the autophagosome functions as a platform facilitating pro-CASP8 activation. Chemoresistance, a common problem in the treatment of cancer, is frequently caused by the downregulation of key mitochondrial death effector proteins. Alternate stress-induced apoptotic pathways, such as the one described here, may become of particular relevance for tackling the problem of chemoresistance in cancer cells.
内质网(ER)应激诱导的细胞死亡通常与线粒体凋亡途径的激活相关,其特征为CYCS(细胞色素c,体细胞)释放、凋亡小体形成和半胱天冬酶激活,从而导致细胞死亡。在本研究中,我们证明,在内质网应激条件下,缺乏CASP9/半胱天冬酶-9或BAX和BAK1且因此线粒体凋亡途径存在缺陷的细胞,仍会经历与半胱天冬酶激活相关的延迟性细胞死亡形式,因此揭示了一种替代性应激诱导的半胱天冬酶激活途径的存在。我们确定CASP8/半胱天冬酶-8为此半胱天冬酶级联反应中的顶端蛋白酶,并发现关键自噬基因ATG5或ATG7中的任何一个基因的敲低都会影响CASP8激活和细胞死亡诱导,突出了自噬在激活这一新型内质网应激诱导的死亡途径中的关键作用。与此一致的是,我们鉴定出一种由ATG5、FADD和前CASP8组成的蛋白质复合物,其组装与半胱天冬酶激活和细胞死亡诱导同时发生。总之,我们的结果揭示了在内质网应激且线粒体凋亡途径存在缺陷的细胞中自噬的毒性潜力,并提出了一种模型,其中自噬体充当促进前CASP8激活的平台。化疗耐药性是癌症治疗中的一个常见问题,通常由关键线粒体死亡效应蛋白的下调引起。替代性应激诱导的凋亡途径,例如此处描述的途径,可能对于解决癌细胞中的化疗耐药性问题特别相关。